军团菌D—RAPD基因分型研究

Genotyping of Legionella Species by D-RAPD

〔目的〕建立军团菌的基因分型方法。〔方法〕采用D—RAPD技术 ,应用 6条简并随机引物对 5株军团菌 (包括 3株嗜肺军团菌 )进行基因分型研究。〔结果〕除引物 4 ,其余 5个随机引物均能对军团菌菌株扩增出特异性的DNA条带 ,其中引物 2的分型效果较好。用引物 2、引物 3两引物 ,引物 2、引物 5两引物 ,引物 3、引物 5两引物 ,分别对 5株军团菌DNA进行扩增 ,结果 3对引物在扩增条带的数量和长度上均能有效区分 5株军团菌。〔结论〕D—RAPD技术快速、简便 ,是在分子水平对致病微生物进行检测、发病机理分析、流行病学研究的理想方法。

To establish a genotyping method of Legionella Species. D RAPD was used to type 5 legionella strains(including 3 legionella pneumophila strains)with 6 degenerate random primers. All the random primers except for primer-4 can amplify the specific fragments of all the legiona strains and the primer-2 is better than other primers to type the legiona strains. 5 legiona strains' DNA were respe ctively amplified by primer-2 and primer-3, primer-2 and primer-5, primer-3 and primer-5 and the result showed that the three pairs of primer3 can well distinguished the 5 legio nella strains in number and length of specific fragments. [Conclusions] D-RAPD is rapid and simple and an ideal method for studies on the detection ,pathogenesis and epidemiology of Legionella Species at molecular level.